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Apr 15, 2015 - School of Pharmacy, Department of Pharmaceutical Chemistry, ITM University, Gwalior ... magnetic susceptibility measurement, LC-MS, H. 1.
WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Sharma et al.

World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 5.210

Volume 4, Issue 06, 696-707.

Research Article

ISSN 2278 – 4357

SYNTHESIS, CHARACTERIZATION AND ANTIMICROBIAL EVALUATION OF COPPER (II) COMPLEX WITH CIPROFLOXACIN ANTIBIOTIC. Richa Kothari1, Brajraj Sharma*2, Swati Sahawal3, Kumari Neha4, Sunanda Kr. Mandal5, Shruti Birthare6 and Vipul Shivhare7 School of Pharmacy, Department of Pharmaceutical Chemistry, ITM University, Gwalior (M.P.) India. ABSTRACT Article Received on 24 March 2015, Revised on 15 April 2015, Accepted on 06 May 2015

New complex of copper (II) with ciprofloxacin & thiosemicarbazide Schiff base has been synthesized, Schiff base ligand and its Cu(II) complex were characterized by various physico -chemical techniques like elemental analysis, m.pt. , TLC, molar conductance measurement,

*Correspondence for

magnetic susceptibility measurement, LC-MS, H1NMR, FT-IR and

Author

electronic spectral studies. The magnetic moment and electronic

Brajraj Sharma

spectra studies suggested distorted octahedral geometry for Cu (II)

School of Pharmacy,

complex. The monoanionic thiosemicarbazonate ligand act in a

Department of Pharmaceutical

tridentate mode, binding through azo-methine nitrogen and sulfur

Chemistry, ITM

atom. The synthesized ligand and its Cu (II) complex were screened

University, Gwalior

for their invitro antibacterial activity using disc diffusion method

(M.P.) India.

against the gram positive and gram negative bacterial strain E. coli and S. aureus using nutrient agar medium. Streptomycin was used as

standard in the test. Both the compounds showed remarkable

antibacterial activity in

comparison to antibiotics. Antioxidant activity of the ligand and its Cu (II) complex was screened using the DPPH scavenging assay. Both the compounds showed potent antioxidant activity in the range 80 – 97%. KEYWORDS: Ciprofloxacin, thiosemicarbazide, spectral studies, antibacterial activity, copper chloride, anti oxidant activity.

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1. INTRODUCTION Ciprofloxacin [1 – cyclopro – pyl – 6 – fluoro – 1,4 – dihydro – 4 – oxo – 7 – (1 – piperaz – inil) – 3 – quinolone carboxylic acid] is a second generation fluoro quinolone that was synthesized for first time in 1987.[1] A well known antibacterial drug with a wide spectrum of activity, it is extremely useful for the treatment of a variety of infections.[2-3] Quinolones are a group of synthetic antibacterial agents now in clinical use already for over thirty years and ciprofloxacin is one of the widely used representatives.[4-5] The interaction of quinolones and metal ions have been thoroughly studied especially due to the interesting biological and chemical properties Ciprofloxacin can usually act as a bidentate ligand through the pyridine nitrogen and one carboxylate oxygen. Literature reveals that metal ions present in complexes accelerate the drug action and the efficacy of the organic therapeutic agents.[6] The pharmacological efficiencies of metal complexes depend on the nature of the metal ions and the ligands.[7] Schiff base lignads based on thiosemicarbazone and their metal complexes have received considerable attention because of their pharmacological properties; they have numerous applications such as antibacterial, antifungal, antioxidant & anticancer agents. [8-10] They can yield more or polynuclear complexes some of which are biological relevant.[11-14] For example, some copper complexes can serve as models for enzymes such as galactose oxidase and may be used as effective oxidants and redox catalysts.[15-16] Drug resistance has become a growing problem in the treatment of infectious diseases caused by bacteria[17] and the discovery and development of effective antibacterial drug with novel mechanisms of action have thus become urgent tasks for infectious disease research programs.[20] The serious medical problem of bacterial and fungal resistance and the rapid rate at which it develops has led to increasing levels of resistance to classical antibiotics.[18-19] Among metal ions of biological importance, the Cu (II) ion is involved in a large number of distorted complexes.[21] Over the past two decades, considerable attention has been paid to metal complexes of Schiff bases containing nitrogen and other donor atoms.[22-23] Bio-organometallic Chemistry is dedicated the study of metallic complexes and their biological applications[24] including the design of new drugs that are more effective than those already known. Thiosemicarbazones are well established as an important class of sulfur donor Schiff base ligands that are particularly useful for transition metal ions. This is due to the remarkable biological activities observed for these compounds, which has been shown to

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be related to their metal complexing ability. As part of our work involving the preparation, characterization and biological evaluation of transition metal complexes, we were interested in obtaining the copper mediated antibiotics from aromatic aldehydes and thisemicarbazide. The main aim of our work to “Synthesize & Characterized the copper mediated antibiotics and then evaluate their antimicrobial and antioxidant activities. Therefore, in the present paper, we report the synthesis, structural, Characterization and Biological evaluation of copper mediated antibiotic derived from ciprofloxacin, O-hydroxybenzaldehyde and CuCl2.2H2O. Synthesized ligand and its Copper complex showed potential antimicrobial and antioxidant activity as compared to ciprofloxacin antibiotic. 2. EXPERIMENTAL 2.1 Chemistry All the glassware’s were dried in an open flame before use in connection with an inert atmosphere. Solvents were evaporated under reduced pressure and evaporation was carried out at temperature C=O of carboxylic group and >C=S bands respectively. All these values are consistent with the tridentate coordination mode of the thiosemicarbazone Schiff base ligand through the thiolate sulfur and azomethine nitrogen atoms.[29] The occurrences of the ν(N-H) band at higher frequencies in the IR spectra of the complexes compared to those observed for the ligands, confirms coordination through the azomethine nitrogen. The ν(C=S) bands at 810 850 cm-1 in the spectra of free ligand. C=N (exo) at 1636 cm-1 and ν C=N (ring) at 1578 cm-1 to be extended 20-40 cm-1 in the spectra of the chelates suggests the involvement of azomethine nitrogen coordination. 3.2 Mass spectral studies The mass spectrum of ligand and its copper complex show its characteristics peaks at m/z values: 222.2, 265, 326.6, 679.2, 402.2, and 645.3 amu. These are in accordance with the proposed structure of ligand and its copper complex. 3.3 H1 NMR Spectral studies The 1HNMR spectra of ligand and its copper complex were obtained in CDCl3 at room temperature using TMS as an internal standard. The aromatic region shows a sharp singlet at δ 7.30 ppm assigned to the phenyl protons and a singlet at δ 2.5 ppm due to methyl protons. The O-H proton of a phenolic group shows a sharp singlet at δ 11.3 ppm. The multiplates observed in the region 6.80-7.92 ppm may be assigned to the aromatic ring proton of the ligand and its copper complex. The 1HNMR spectra of the copper complex shows signals corresponding to –CH3, -NH2, NH and –OH protons at 2025 (S, 3H), 7.40 – 7.43 (m, 3H), 8.1-8.9 (2H), 10.0 (S, 1H) and 11.4 (S, 1H) respectively. The 1HNMR spectrum of the chelates confirms the imino –NH group in the coordination with copper ions. Some hydrogen atoms values of δ were not observed precisely due to overlapping with the signals of the aromatic hydrogen atoms of the ligand and copper chelate. 1HNMR integration and signal multiplicity are in agreement with the proposed structures. In the 1HNMR spectra of the copper complex a high frequency shift appeared at 2.2 ppm, for the methyl hydrogen atoms (C-CH3) compared to the spectra of the ligand and evidences the coordination through the azomethine nitrogen atoms.

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3.4 Physiological activity The culture medium was prepared by the addition of 1.5 g of Beef extract, 1.5 g of yeast extract, 5 g of peptone, 1g of dextrose, 3.5 g of NaCl and 3.68 g of dipotassium hydrogen phosphate, 1.32 g of monopotassium dihydrogen phosphate and 15 g of agar in one liter of water. The pH of the medium was adjusted to 7.0. The medium is sterilized at 121 °C in an autoclave at 15 lbs pressure for 15 – 20 min. The medium was cooled to 45 – 50 °C. 20 ml of the medium was poured in each of the Petri-dishes. All the apparatus were sterilized before use E. Coli and S. aureus were introduced in to the Petri – plates and were grown in incubator at 37°C for 24 hrs. 3.5 Bacteriological Testing Bacteriological testing was done using disc diffusion method. Paper discs were soaked in the DMF solutions of the ligand and its copper complex. Solutions were prepared in four different concentrations i.e. 100 µg/ml, 200 µg/ml, 300 µg/ml and 400 µg/ml in DMSO. Paper discs were soaked in these solutions. They were dried. The dried paper discs were introduced in to the completely grown culture plates of E. coli and S. aureus and then incubate the plates at 37 °C for 24 hrs. It was found that all the metal complexes were active against test bacteria as compared ligands. Table 3: Bactericidal activity of the ligand and its metal complexes after 24 hrs. S.No

Ligand / Complex

1. 2. 3

C18H24N6O2FS (Ligand) [Cu(C36H48N12F2S2)]Cl2 Streptomycin Standard drug (C17 H18 F N3 O3)

Zone of inhibition after 24 hrs in concentration µg/ml Escherichia Coli S. Aureus 100 200 300 400 100 200 300 400 µg/ µg/ µg/ µg/ µg/ml µg/ml µg/ml µg/ ml ml ml ml ml 8.8 10.5 12.5 14.2 9.5 12 13.5 15.5 15 18 20 22 16 18 20 24 26 32 34 38 24 28 33 40

The high antibacterial activity of copper complex may be due to coordination and chelation which tend to make copper complex act as powerful and potent bacteriostatic agents, thus inhibiting the growth of the bacteria. In copper complex, the positive charge on the copper is partially shared with the donor atoms present in the ligand and there may be Π e delocalization over the whole chelate. The increased activity of the copper chelate can be explained on the basis of chelation theory. On chelation, the polarity of the copper ion is reduced largely due to the overlap of the ligand orbital and the partial sharing of the positive

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charge of the copper ion with the donor groups. This increases the lipophilic character of copper complex seem to be the reason of their enhanced potent antibacterial activity. There are some other factors is also increase the activity such as solubility, conductivity and bond length between the copper and ligand. Copper complex was active due to the presence of imino group in the complexes. 3.6 Antioxidant activity The RSA of Schiff base ligand and its copper complex may be due to the presence of >NH groups which may donate an electron or hydrogen atom to DPPH and form a stable free radical. This radical can be stabilized by delocalization. This fact confirm their good electron and / a hydrogen atom donating ability and to act as free radical scavenger. However, none of compound exhibited radical scavenging activity better than standard ascorbic acid. Table 4: Antioxidant activity of ligand and its copper complex. S.No Complex

1 2 5

C18H24N6O2FS [Cu(C36H48N12F2S2)]Cl2 Ascorbic acid

Percentage scavenging activity of ligand and its copper (II) complex. 200 μɡ 400 600μɡ/ml 800μɡ/ml 1000μɡ /ml μɡ/ml /ml 81.8 80 89.6 90.2 93 82 84 90 92.8 94 84 86 92 93.8 96

Schiff base ligand and its copper complex showed significant free radical scavenging action against DPPH induced release of free radicals at different concentration 200 µg/ml, 400 µg/ml, 600 µg/ml, 800 µg/ml, 1000 µg/ml. Ascorbic acid is used as a standard. 3.7 CONCLUSION This paper presents “Synthesis, Characterization and Antimicrobial Evaluation of Copper (II) Complex with Ciprofloxacin Antibiotic”. Both the compounds was synthesized and well Characterized in detail by FT – IR, H1NMR, LC-MS, Electronic spectra, Elemental analysis, melting point, TLC and molar conductance data. Spectroscopic studies suggested a six coordinated octahedral geometry for Cu (II) complex. Conductivity measurement supported the non-electrolytic nature of Cu (II) complex. IR spectra indicate that the Schiff base ligand acts in tridentate fashion by bonding to the copper ion through the imine nitrogen and oxygen atom. The mass spectra show the monomeric nature of the complex. Antibacterial results show that the copper (II) complex exhibited good antibacterial potential. Antioxidant activity results show that the ligands and its copper (II) complex exhibited good antioxidant activity. www.wjpps.com

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The data reported in this paper may be helpful guide for the medicinal chemist who is working in this area. 3.8 ACKNOWLEDGEMENTS The authors are thankful to the Chancellor, Vice Chancellor, Managing Director, ITM University, Gwalior, ITM University, Gwalior, for their support and co-operation. RK is grateful to MPCST for providing financial assistance in the form of grant no. (Council order No. 4566/ Cst/ R&D/2010). REFERENCES 1. W. Mertindale, “Martindale – the extra Pharmacopeia”, 30th Edition, the Pharmaceutical Press, London, 1993. 2. P. Ball, “The quinolones, Academic Press, San Diego, 2000. 3. G. Sheehan, N. S. Y. Chew, “The history of quinolones” In; A.R. Ronald, D.E. Low, Eds., Fluoroquinolone Antibiotics, Birkhauser, Basel: 2003, pp. 1-10. 4. I. Turel, “The Interactions of metal ions with quinolone Antibacterial Agents”, Coordination Chemistry Reviews, 232(1-2): 27-47. 5. B. E. Scully, M. Nakatomi, C. Ores, S. Davidson, H. C. Neu, “Ciprofloxacin therapy in cycstic fibrosis”, The american Journal of Medicin, 1987; 82(4A): 196-201. 6. Z. A. Siddiqui, M. Khalid, S. Kumar, M. Shalird and S. Noor, “Antimicrobial and SOD activities of Novel Transition Metal Complexes of pyridine – 2, 6 – dicarboxylic acid containing 4 – picoline as Auxiliary ligand”, European Journal of Medicinal Chemistry, 2010; 45(1): 264-269. 7. S. Delaney, M. Pascaly, P.K. Bhattacharya, K. Han and J.K. Barton, “Oxidative Damage by Ruthenium complexes containing the dipyridophenazine ligand or its derivatives, A focus on Intercalation, Inorganic Chemistry, 2002; 41(7): 1966-1974. 8. M. B. Ferraria, F. Bisceglie, A. Buschini, S. Franzoni, G. Pelosi, S. Pinelli, P. Taraconi, M. Tavone, Synthesis, structural characterization and antiproliferative and toxic bioactivities

of

copper

(II)

and

nickel

(II)

citronellal

N-4

ethylmorpholine

thiosemicarbazonates, J. Inorg. Biochem. 2010; 104:199. 9. D. X. West, E. Liberta, S. B. Padhyl, R. C. Chikate, P. B. Sonawane, A. S. Kumbar, R. S. Yerands, Macrocyclization of the cyclic thiosemicarbazones with mercury salts; coordination Chem. Rev, 1993; 123: 49.

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10. D. X. West, S.B. Padhye, P. B. Sonawane, Structural and Physical correlation in the biological properties of transition metal N-heterocyclic thiosemicarbazones and Salkyldithiocarbazate, Struct. Bond 76, 19, 1-50. 11. Y. Haidue, A. Silverstru, Coord. Chem. Rev.,1990; 99: 253. 12. O. E. Ichiro, D. Busch, H. Shull, Bioinorganic Chemistry, Allym and Bacon, Boston, MA, 1977. 13. R.W. Hay, J. R. Dilworth, K. B. Nolan, Bioinorganic Chemistry, vol. 1, Jai Press, London 1991. 14. V. W. W. Yam, Y. L. Pui, W.P. Li, K.K.W.Lo, K.K. Cheung, J. Chem. Soc. Dalton Trans. 1998; 3615. 15. M.R. Malachowshi, B.T. Dorsey, M.J. Parker, M. E. Adams, R. S. Kelly, “Probing the catalytic properties of copper (II) complexes of appended cyclams: correlations between catalysis and stability constants or electro chemical properties” Polyhedron, 17, 1998, 1289. 16. B. J. Hathaway, G. Wilkinson, R. Gillard, J. A. McCleverty(Eds.) , Comprehension Coordination Chemistry, Pergamon, Oxford, 1987. 17. N. Raman, A. Sakthivel, K. Rajasekaran, Synthesis and spectral characterization of antifungal sensitive Schiff base transition metal complexes. Mycobiology, 2007; 35: 150153. 18. S. A. Rice, M. Givskov, P. Steinberg, S. Kjellsberg, Bacterial signals and antagonists: The interaction between bacteria and higher organism, J. Mol. Microbiol. Biotechnol., 1999; 23-31. 19. A. Ironmonger, B. Whittaker, J. Andrew, B. Baron, J. Chris, E. Alison, G. Ashcroft, A. Nelson, Scanning conformational space with a library of stereo and region-chemically diverse aminoglycoside derivatives. The discovery of new ligands for RNA sequences. Org. Biomol. Chem, 2007; 5: 1081-1086. 20. G. Kumar, D. Kumar, S. Devi, R. Verma, R. Johari; Synthesis, Spectral characterization of biologically active compounds derived from oxalyldihydrazide and 5- tert – butyl -2 – hydroxyl – 3 – (3 – Phenylpent – 3 – yl) benzaldehyde and their Cu(II), Ni(II) and Co(II) complexes. Int. J. Eng. Sci. Technol, 2011; 3: 1630-1635. 21. K. Singh, M. S. Barwa, P. Tyagi “Synthesis, Characterization and biological studies of Co(II), Ni(II), Cu(II) and Zn(II) complexes with bidentate Schiff bases derived by heterocyclic ketone” European Journal of Medicinal Chemisrty, 2006; 41(1): 147-153.

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22. J. A. Obaleye, J. F. Adediji, M. A. Adebayo, “Synthesis and biological activities on metal complexes of 2,5 – diamino – 1,3,4 – thiadiazole derived from semicarbazide hydrochloride “, Molecules, 2011; 16(7): 5861-5874. 23. X. Tai, X. Yin, Q. Chem, M. Tan, Synthesis of some transition metal complexes of a novel Schiff base ligand derived from 2,2 – bis(p – methoxyphenylamine) and salicylaldehyde, Molecules, 2003; 8(5): 439-443. 24. R. Kothari, B. Sharma “Template synthesis and spectral characterization of hexa aza macrocyclic Co(II), Ni(II) and Cu(II) complexes: Antimicrobial and DNA Cleavage studies”, Int. J. Chem, 2013; 2(2): 199-208. 25. Prasad R. N., Gupta S. and Jangir B., J. Indian Chem. Soc., 2007; 84: 1191-1194. 26. A.A. Al-Amiery, A. Mohammed, H. Ibrahim, A. Abbas, Study the biological activities of Tribulus terrestris extracts, World Acad. Sci. Eng. Technol. 2009; 57: 433-435. 27. Prasad, R.N. ; Mathur, M.; Upadhayay, A. J. Indian Chem. Soc. 2007; 84: 1202. 28. Roy, 2009, Naresh Kumar and Ramesh, 2004. 29. D.R. Klayman, J.F. Bartoseich , T. S. Griffin, C. J. Manson, J.P. Scovill, Pesticidal activities of some Schiff bases derived from Benzoin, Salicylaldehyde, Aminophenol and 2, 4- dinitrophenyl hydrazine J.Med. Chem. 1979; 22: 855.

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